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Volume 149, Issue 2

An amino acid change near the carboxyl terminus of the regulatory protein Rgg affects its abilities to bind DNA and influence expression of the glucosyltransferase gene Free

Abstract

The glucosyltransferase structural gene, , is located immediately downstream from its positive transcriptional regulatory determinant, . Recent genetic studies have indicated that the 3′ end of is involved either directly as a binding site or indirectly, e.g. by playing a role in secondary structure, in the interaction of Rgg with the promoter. A previously identified spontaneous mutant with a point mutation near the 3′ end of had only ∼25 % of the parental level of glucosyltransferase activity. To determine if this decreased activity was due to a change in the DNA binding site of -acting Rgg, or due to a change in the Rgg protein itself, complementation analyses and DNA-binding studies were performed. In Rgg-deficient strains, the chromosomal point mutation did not influence the ability of plasmid-borne to increase glucosyltransferase expression. However, plasmids carrying parental were able to increase glucosyltransferase activity and expression of a promoter fusion to a greater extent than plasmids carrying the mutant allele, indicating that the mutant Rgg protein had decreased activity. The ability of NH-terminal (hexahistidine) tagged proteins to bind to a 107 bp dsDNA fragment corresponding to the region immediately upstream of was demonstrated by surface plasmon resonance. Despite their differences in activity, both mutant and parental recombinant Rgg proteins bound to this dsDNA, albeit with different strengths. These studies provide insights into functional domains of Rgg which influence glucosyltransferase expression, and may have implications for Rgg-like regulatory proteins in related bacteria.

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  • Accepted:
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  • Published Online:
Keyword(s): GTF, glucosyltransferase , Spp, sucrose-promoted phenotype and SPR, surface plasmon resonance
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2003-02-01
2024-04-27
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An amino acid change near the carboxyl terminus of the Streptococcus gordonii regulatory protein Rgg affects its abilities to bind DNA and influence expression of the glucosyltransferase gene gtfG
Microbiology 149, 399 (2003); https://doi.org/10.1099/mic.0.25983-0
/content/journal/micro/10.1099/mic.0.25983-0
/content/journal/micro/10.1099/mic.0.25983-0
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